Aedes albopictus (Skuse) (Diptera: Culicidae) and Aedes aegypti (L.) (Diptera: Culicidae) are invasive container mosquitoes that are of potential medical importance in the southern United States. Seeds (acorns) and leaves from oak trees can contribute seasonally to the detritus of larval container habitats. Herein, we examined the effect of acorns and leaves from the southern live oak (Quercus virginiana Mill.), which has a concomitant range with these mosquitoes, on the population performance of Ae. aegypti and Ae. albopictus. Three levels of acorns and oak leaves were used (0.99, 1.98, and 7.94 g), along with two mixtures (leaf + acorn: 0.50 + 1.48 and 1.48 + 0.50 g). Tannins, secondary plant metabolites that effect herbivory, were measured across all treatment levels; nitrogen and carbon was also measured for detritus and representative females. Survival, female mass, development time, and λ (per capita rate of population increase) were used to evaluate population performance of both species. Detritus amount but not type led to differences in tannins; however, these differences did not correspond to differences in performance. Acorns had higher carbon and C:N than leaves. Survival for both species was lower in medium amounts of acorns. Female mass varied with leaf amount, whereas development time differed between amounts of pure leaf and acorn. λ was lowest in medium and high acorns compared with leaves or mixtures. Thus, acorns do appear to limit mosquito survival and affect population growth, suggesting that inputs of this common detritus type may negatively affect container Aedes production.

Vertical habitat stratification in populations of Aedes hendersoni Cockerell (Diptera: Culicidae) and Aedes triseriatus (Say) (Diptera: Culicidae) has been observed to varying degrees throughout the lspecies' sympatric range, and potential causes of the phenomenon, including species competition and interaction, have been debated extensively. Stratification patterns in oviposition in allopatric, sympatric, and marginally sympatric populations of both species were investigated and compared in this research to detect any pattern differences related to species composition. Expected patterns were observed in sympatric populations, with Ae. hendersoni preferentially ovipositing in canopy habitats, whereas Ae. triseriatus preferred basal habitats. Allopatric populations presented a strong shift toward basal preference in the former and a slighter but significant shift toward canopy in the latter. Marginal populations of Ae. hendersoni showed intermediate height preferences, whereas preferences of marginal and sympatric Ae. triseriatus did not differ. The convergence of habitat selection in allopatric populations and corresponding divergence in sympatric populations support interspecific competition-based hypotheses regarding the origin of the stratification phenomenon, although plausible alternative or contributing explanations are numerous and warrant further study.

We tested the effect of the native container-dwelling predator Toxorhynchites rutilus on the codominant container-dwelling mosquitoes: native Aedes triseriatus and invasive Aedes japonicus. We established two predator treatments (predator, no predator) by removing T. rutilus from all containers, and stocking T. rutilus larvae (1/3.5 liters) in the predator treatment. Removal and stocking was repeated every 3 d and established significantly different predator abundances in both large and small containers. Repeated-measures analysis of variance (ANOVA) on standard samples showed larvae+pupae/liter of A. japonicus was greater without versus with predation, and this difference increased across samples. In contrast, repeated-measures ANOVA showed larvae+pupae/liter of A. triseriatus was statistically indistinguishable for predation treatments and was greater in small versus large containers.Thus, predation reduced invasive A. japonicus while having no detectable effect on A. triseriatus larvae and pupae. A final destructive census of pupae showed that predation reduced pupae/liter of both species, but this effect was greater and more consistent across container sizes for A. japonicus. Predator effects on abundances were not products of the nonlethal effect of predator avoidance by ovipositing females, as T. rutilus presence did not lead to reduced egg inputs by either Aedes, nor by Aedes spp. as a group. Effects of predation thus are best explained by differential success of developing larvae due to the greater lethal effect of T. rutilus on A. japonicus than on A. triseriatus.Thus, this system is consistent with the hypothesis that native predators can limit success and potential impacts of invasive mosquitoes.

In several areas where Aedes aegypti (L.) (Diptera: Culicidae) and Aedes albopictus (Skuse) (Diptera: Culicidae) have come in contact following successful invasions, Ae. aegypti have been rapidly displaced by Ae. albopictus. Recent work has confirmed that mating interference, in the form of satyrization, is likely a driving factor in these competitive displacements. However, in sites of sympatry, Ae. aegypti females evolve resistance to satyrization, and in the laboratory, satyrization-susceptible Ae. aegypti can evolve resistance within a few generations of cage exposure to Ae. albopictus. The resistance trait may be costly to maintain, as satyrization-resistant females have been shown to invest more time in mate selection. Here, we show that increased satyrization-resistance does not persist in the absence of satyrization pressure, confirming a cost to maintaining this trait. Ae. aegypti lines that had been previously selected for increased satyrization-resistance demonstrated rapid decreases in this trait over eight generations. Support for the hypothesis that condition-dependent mate preferences lead to variation in mating choice among individuals of differing quality within a population has been accumulating. Here we examine how age-related changes in reproductive effort or reproductive value may result in age-dependent mate choice. We hypothesize that older, lower quality individuals may be more likely to accept a subpar mating, in this case an interspecific mating. Our results demonstrate that mosquito age significantly affects interspecific mating rates with older mosquitoes (male and female) engaging in interspecific mating more frequently than younger counterparts (26.32 vs 9.41%). We discuss the possibility of age-related signal variation causing the breakdown of mating barriers with age.

The mosquitoes Aedes aegypti (L.) (Diptera: Culicidae) and Aedes albopictus (Skuse) (Diptera: Culicidae) are ecologically similar species that have evolved independently in their native ranges, Ae. aegypti in Africa and Ae. albopictus in Asia. Where their invasive ranges overlap, interspecific mating is facilitated by both species swarming to mate around bloodmeal hosts during daylight. Here, we test hypotheses to account for variation in spermathecal filling in females of Ae. aegypti and Ae. albopictus, particularly, the effect of mosquito size on the number of spermathecae containing sperm after intraspecific and interspecifc matings of these two species. Results show that significantly more spermathecae contained sperm in large than in small Ae. albopictus females, but there was no effect of Ae. aegypti female body size on the average number of spermathecae filled. Among inseminated females, there was no effect of male size or cross-type on number of spermathecae with sperm. Differences in the effect of female size on the number of spermathecae that store sperm in Ae. aegypti and Ae. albopictus imply differences in the mating biology of these two species.

Zika virus (ZIKV) first spread into Brazil in 2013 and is now present throughout the Americas. In 2016, Florida witnessed the beginnings of local ZIKV transmission. No reports of local transmission have been reported for 2018; however, travel related cases continue to be reported. Recurrence of local transmission in the United States is a major public health risk in Florida where vectors Aedes aegypti (Diptera: Culicidae, Linnaeus) and Aedes albopictus (Diptera: Culicidae, Skuse) are abundant and there is a high potential for virus reintroduction. A dose-response study was used to evaluate susceptibility and transmission potential of Florida Ae. aegypti and Ae. albopictus to ZIKV originating from Puerto Rico. Mosquitoes were orally exposed to one of three doses of ZIKV. Higher doses of infected blood resulted in overall greater infection rates in both mosquito species. Ae. aegypti and Ae. albopictus were susceptible to infection with ZIKV and revealed a significant species by dose interaction. At low doses, Ae. aegypti was significantly less susceptible to infection with ZIKV than Ae. albopictus (6.7% and 44.4%, respectively). In contrast, at high doses, Ae. aegypti was significantly more susceptible to infection than Ae. albopictus (75.8% and 53.8%, respectively). No significant differences were observed between Ae. aegypti and Ae. albopictus in disseminated infection (0–75%) and saliva infection (0–52.4%). These observations suggest greater susceptibility to infection for Ae. albopictus at lower doses likely encountered by viremic humans. However, low disseminated infection and saliva infection for Ae. albopictus, combined with catholic feeding behavior, are likely to limit transmission potential relative to Ae. aegypti.

House dust mites (HDMs) are the major constituents of house dust (HD). HD and HDM sensitization is well documented worldwide. Storage mite (SM) sensitization is presently lacking from India.The present study evaluated the sensitization of both HDM and SM among 372 allergic rhinitis patients reported to the Allergy and Asthma Research Center of Kolkata metropolitan, India. HD samples were collected from the patients' home and analyzed for the major constituent mites. HD and six constituent mites Dermatophagoides pteronyssinus (DP), Dermatophagoides farina Hughes (Acari: Pyroglyphidae) (DF), Blomia tropicalis (BT) Bronswijk et al., Acarus siro Linnaeus (Acari: Acaridae) (AS), Lepidoglyphus destructor (Schrank) (LD), and Tyrophagus putrescentiae (Schrank) (Acari: Acaridae) (TP) are tested for the allergenic potential through Skin Prick Test (SPT). Three SMs, namely AS, LD, and TP, were newly included in the mite SPT extract for the first time in Kolkata. In total, 330 patients showed significant positive SPT toward any one allergen tested. HD was the major elicitor exhibiting 92.42% response. Individuals of age group 15-40 were the worst sufferers. DF showed the highest sensitization (87.87%) among the dust mites. The SMs also contributed significantly to prove their sensitizing potential. SPT rates for AS, LD, and TP were 33, 25, and 18%, respectively. SPT grades and total Immunoglobulin E (IgE) were positively correlated for each of the allergens. Most of the patients were multi-sensitized (95%) and represented markedly high total IgE levels (>500 IU/ml). Three SMs proved to be significant allergens for the studied population.The sensitization toward these SMs is first time reported from India and can be recommended for inclusion of routine SPT for better outcome in the future.

Biting midges (Ceratopogonidae: Culicoides) are vectors of bluetongue virus and epizootic hemorrhagic disease virus which cause significant morbidity and mortality in ruminants. Recently, ultraviolet light emitting diodes (UV/ LEDs) in conjunction with suction traps have been widely utilized for Culicoides spp. collections. Despite the use of these traps, limited work has been done comparing sampling variability associated with these light types with and without CO₂. For this objective, mini-CDC light traps with four different attractant combinations were operated at eight sites across Florida between April and October 2017. Trap attractants included white-incandescent lights and UV/LEDs with and without CO₂ to determine optimum combinations of light type and attractant for species richness, diversity, and abundance of Culicoides spp. in Florida. The results of the study demonstrate that traps with UV/LED light collect greater richness, diversity, and abundance of Culicoides species than traps with white-incandescent light. Addition of CO₂ resulted in greater diversity in traps with UV/LED lights, but lower diversity in traps with white-incandescent light. Therefore, CO₂ may be used to increase the abundance of Culicoides spp. collected by traps, regardless of light type, but the ability for CO₂ to attract a higher number and diversity of species to traps varies by the light type used. Therefore, we suggest using CO₂ primarily in conjunction with UV/LED light. When CO₂ is not available, UV/LED light used alone can be substituted without a significant loss in species richness or diversity, although abundance of most Culicoides species will be significantly lower in the absence of CO₂.

Bed bugs (Cimex lectularius L.) (Hemiptera: Cimicidae) are obligate hematophagous ectoparasites, and, therefore, must locate suitable hosts to ensure survival and reproduction. Their largely nocturnal activity suggests that chemosensory and thermosensory cues would play critical roles in host location. Yet, the importance of olfaction in host attraction of bed bugs remains unclear. We developed and validated a Y-tube, two-choice olfactometer and tested its suitability for investigating attraction to human odors (from skin swabs). Olfactometer orientation significantly affected the percentage of bed bugs that were activated by human odors, with significantly more bed bugs responding when the olfactometer was oriented vertically (bug introduced at bottom of the olfactometer) compared with all other orientations. Starved (7–10 d) adult males, mated females, and nymphs responded (47–77% moved up the olfactometer and made a choice) when human odors were present in the olfactometer, while starved, unmated females did not respond. Skin swabs from all five human participants elicited high response rates (65–82%), and bed bugs from four different populations responded to skin swabs (40–82% response rate). However, in all assays including those resulting in relatively low response rates, bed bugs exhibited >90% preference for human odors over blank controls. These results provide strong evidence that bed bugs can respond and orient towards human odors, independently of all other host cues. Furthermore, the validated olfactometer should enable rapid and efficient evaluations of bed bug behavioral responses to semiochemicals.

Nasonia vitripennis (Walker) (Hymenoptera: Pteromalidae) is a common ectoparasitoid that attacks the puparia of fly species of forensic importance, including blow flies, flesh flies, and house flies. The developmental time of N. vitripennis can be added to the host development time, providing the potential for an extended minimum postmortem interval timeframe in cases where traditional dipteran larval forensic timeline indicators have completed their development. In this study, we used Boettcherisca peregrina Robineau-Desvoidy (Diptera: Sarcophagidae) as the host for N. vitripennis and studied the development of N. vitripennis at seven constant temperatures between 16 and 34°C.The developmental process was divided into 12 periods, and we measured developmental time, changes in larval body length, and thermal requirements for development, including developmental thresholds and thermal constant. Using these data, we created an isomorphen diagram and thermal summation model of N. vitripennis. The total developmental duration of parasitoids from egg to adult emergence at 16, 19, 22, 25, 28 and 31°C were 953.3 ± 20.0, 698.0 ± 7.7, 508.7 ± 13.6, 354.7 ± 4.6, 272.0 ± 13.9, and 232.0 ± 11.3 h, respectively. Nasonia vitripennis did not emerge at 34°C. The developmental threshold temperature of N. vitripennis was 11.52°C, and the thermal summation constant was 4768.8 degree hours.

The study of the arthropod fauna involved in carcass decomposition is crucial for medicolegal forensic purposes. This knowledge is especially needed in countries with high levels of criminality and where the arthropod fauna is still under-recorded and not well known, such as Angola. We present data from the first survey on sarcosaprophagous flies and beetles in Angola, conducted in Luanda over a 17-d period, in the wet summer season. In total, 881 adult and immature specimens, belonging to 18 species, were collected from three pig carcasses that were sampled daily. Three species were not previously known from Angola. The dominant flies were Chrysomya albiceps (Wiedemann, 1819) (Diptera: Calliphoridae) and Chrysomya megacephala (Fabricius, 1794), but Chrysomya marginalis (Wiedemann, 1830), Chrysomya putoria (Wiedemann, 1830), and Liosarcophaga emmrichiana (Lehrer, 2002) (Diptera: Sarcophagidae) also bred on the carcasses. The dominant beetles were Dermestes maculatus De Geer, 1774 (Coleoptera: Dermestidae), Saprinus splendens (Paykull 1811) (Coleoptera: Histeridae), and Necrobia rufipes (De Geer, 1775) (Coleoptera: Cleridae). Because of the hot weather during the experimental period (∼27°C), decomposition was extremely fast. Periods of insect activity on carcasses are presented and comparisons are made with other carrion communities reported from the Afrotropical region.

Asian longhorned or bush tick (Haemaphysalis longicornis Neumann) is a vector species of considerable medical and veterinary importance within its native range in East Asia, and in introduced areas of Australia and Oceania. Recently, this tick species was detected in several regions of the United States. This study aimed at modeling areas suitable for H. longicornis in North America using maximum entropy distribution modeling or Maxent. Occurrence records of H. longicornis within its present range were obtained from published literature. The Maxent model contained a small number of a priori climatic and ecological variables. Annual temperature, precipitation, and ecological zones were found the most important in creating sensitive and specific model (success rate = 91.8%) that had a good fit to the existing data. The model predicted suitable H. longicornis habitat in most of eastern North America from southern Canada to the Gulf Coast, and in a small temperate area on the West Coast. Coastal areas were among the highest ranked suitable habitat. Another highly suitable region was identified in mid-western and southern United States where Heartland virus transmission takes place. This finding is of concern, since H. longicornis ticks vector a closely related virus in East Asia. Delineation of areas suitable for H. longicornis can facilitate detection, prepare public health authorities, and inform the general public about this potentially pestiferous and medically important species.

The flesh flies (Diptera: Sarcophagidae) are significant in forensic investigations. The mitochondrial genome (mitogeome) has been widely used as genetic markers for phylogenetic analysis and species identification. To further understand the mitogenome-level features in Sarcophagidae, the complete mitogenome of Sarcophaga formosensis (Kirneret Lopes, 1961) (Diptera: Sarcophagidae) and Sarcophaga misera (Walker, 1849) (Diptera: Sarcophagidae) was firstly sequenced, annotated, and compared with other 13 Sarcophagidae species. The result indicated that the gene arrangement, gene content, base composition, and codon usage were conserved in the ancestral arthropod. Evolutionary rate of the mitogenome fragments revealed that the nonsynonymous and synonymous substitution rates (Ka and Ks) ratio was less than 1.00, indicating these variable sites under strong purifying selection. Almost all transfer RNA genes (tRNAs) have typical clover-leaf structures within these sarcophagid mitogenomes, except tRNA-Ser (AGN) is lack of the dihydrouridine arm. This comparative mitogenomic analysis sheds light on the architecture and evolution of mitogenomes in the Sarcophagidae. Phylogenetic analyses containing the interspecific distances from different regions in these species provided us new insights into the application of these effective genetic markers for species identification of flesh flies.

A man-biting black fly species so far regarded as Simulium rufibasis Brunetti in Thailand was morphologically analyzed. It was found to be almost indistinguishable in the female from S. rufibasis sensu stricto but distinguishable in the male by the slender fore basitarsus and abdominal segments 2, 6, and 7 each with a pair of shiny dorsolateral patches, and in the pupa by the absence of spine-combs on abdominal segment 7 and terminal hooks on segment 9. It is described as a new species, Simulium tenebrosum. Taxonomic notes are given to separate it from all 11 known species of the S. rufibasis subgroup of the Simulium (Simulium) tuberosum species-group. An analysis of the COI gene sequences shows that this new species is distantly separated from the closely related species.

Trichophoromyia iorlandobaratai sp. n. Vasconcelos dos Santos, Santos Neto, Sánchez Uzcátegui & Galardo is described and illustrated on the basis of male specimens collected in the municipality of Itaituba, Pará State, Brazilian Amazonia.This species can be differentiated from other morphologically similar species within Trichophoromyia by means of the male characters such as the ratio between the aedeagal ducts/sperm pump, shape and distribution of setae in the gonocoxite, and particularly the distinct shape of paramere.

The mosquito midgut is divided into two regions named anterior midgut (AMG) and posterior midgut (PMG). The midgut expands intensely after the blood ingestion to accommodate a large amount of ingested food.To efficiently support the bloodmeal-induced changes, the organization of the visceral muscle fibers has significant adjustments. This study describes the spatial organization of the Anopheles aquasalis (Culicidae, Anophelinae) midgut muscle network and morphological changes after bloodmeal ingestion and infection with Plasmodium vivax (Haemosporida, Plasmodiidae). The midgut muscle network is composed of two types of fibers: longitudinal and circular. The two types of muscle fibers are composed of thick and thin filaments, similar to myosin and actin, respectively. Invagination of sarcoplasm membrane forms the T-system tubules. Sarcoplasmic reticulum cisternae have been observed in association with these invaginations. At different times after the bloodmeal, the fibers in the AMG are not modified. A remarkable dilation characterizes the transitional area between the AMG and the PMG. In the PMG surface, after the completion of bloodmeal ingestion, the stretched muscle fibers became discontinued. At 72 h after bloodmeal digestion, it is possible to observe the presence of disorganized muscle fibers in the midgut regions. The Plasmodium oocyst development along the basal layer of the midgut does not have a significant role in the visceral musculature distribution. This study provides features of the visceral musculature at different blood feeding times of An. aquasalis and shows important changes in midgut topography including when the mosquitoes are infected with P. vivax.

Simulium (Gomphostilbia) yvonneae sp. nov. is described based on adults, pupae, and mature larvae from Vietnam. This new species belongs to the Simulium duolongum subgroup in the S. batoense species-group of the subgenus Gomphostilbia Enderlein. It is distinguished by having a relatively larger number of male upper-eye facets in 16 vertical columns and 16 horizontal rows and a pupal gill with eight filaments arranged as 3+(1+2)+2 from dorsal to ventral, of which two filaments of the ventral pair are 1.8 times as long as the longest filament of the middle and dorsal triplets. Morphological comparisons are made to distinguish this new species from all 22 related species. The genetic distinctiveness of this new species in the S. duolongum subgroup is also presented based on the DNA barcoding COI gene.

Making optimal oviposition decisions is especially important for female carrion colonizing insects whose larvae often depend on ephemeral resources. Optimal oviposition theory predicts that females should exhibit behaviors that will maximize the performance of their offspring. Oviposition can be influenced by a variety of factors, including temperature. The aim of this study was to investigate the effects of temperature on the oviposition behavior and egg load of two blow fly species native to southern Ontario: Phormia regina Meigen, and Lucilia sericata Meigen (Diptera: Calliphoridae). Using fetal pig carcasses as an oviposition substrate, we assessed the length of time until the first oviposition event, preferred oviposition sites, and the total number of eggs oviposited. For each of the two species, five different temperature treatments ranging from 15 to 35°C were used. Temperature influenced time to the first oviposition event, as well as the number of eggs deposited by L. sericata and P. regina. As temperature increased, oviposition occurred faster with more eggs deposited for these two species. Female P. regina, in particular, demonstrated a higher egg load with increasing temperature. Differences in optimal temperatures for reproduction, including egg load and oviposition, may be a mechanism driving seasonal temporal resource partitioning leading to species coexistence utilizing ephemeral resources.

The numbers of Ixodes ricinus (L.) and Dermacentor reticulatus (Fabricius) larvae and nymphs attached to small mammals are influenced by interspecific competition.The present study analyses data collected over several years in two study areas: Kosewo Górne (Mazurian District, N Poland; between July 1997 and July 2009) and Białowieża Primeval Forest (E Poland; in July 2007). In total, 975 ticks were collected from striped field mice (Apodemus agrarius), yellow-necked mice (A. flavicollis), bank voles (Myodes glareolus), and root voles (Microtus oeconomus). In total, of the 203 investigated rodents, 137 were infested with ticks and 39 demonstrated mixed infection. The numbers of the two tick species found on Apodemus mice were significantly negatively correlated with those on root voles; similarly, although bank voles were significantly more frequently infested by I. ricinus than by D. reticulatus, the reverse was observed in root voles. In addition, among the voles, each tick species was found in different locations on the host body, which could also result from competitive interactions; furthermore, competitive release regarding microhabitat selection was observed on hosts infested with one tick species.This competition may be driven by the limited area of host body available for foraging by ticks, i.e., safe areas of high vascularization covered by thin skin. However, the mechanisms of such competition require further investigation.

The composition, density, diversity, and temporal distribution of mosquito species and the influence of temperature, relative humidity, and rainfall on these data were investigated in 50 sites across five land cover classes (forest, savannah, barren, village, and agriculture) in southeastern Senegal. Mosquitoes were collected monthly in each site between June 2009 and March 2011, with three people collecting mosquitoes landing on their legs for one to four consecutive days. In total, 81,219 specimens, belonging to 60 species and 7 genera, were collected. The most abundant species were Aedes furcifer (Edwards) (Diptera: Culicidae) (20.7%), Ae. vittatus (Bigot) (19.5%), Ae. dalzieli (Theobald) (14.7%), and Ae. luteocephalus (Newstead) (13.7%). Ae. dalzieli, Ae. furcifer, Ae. vittatus, Ae. luteocephalus, Ae. taylori Edwards, Ae. africanus (Theobald), Ae. minutus (Theobald), Anopheles coustani Laveran, Culex quinquefasciatus Say, and Mansonia uniformis (Theobald) comprised ≥10% of the total collection, in at least one land cover. The lowest species richness and Brillouin diversity index (HB = 1.55) were observed in the forest-canopy.The urban-indoor fauna showed the highest dissimilarity with other land covers and was most similar to the urban-outdoor fauna following Jaccard and Morisita index. Mosquito abundance peaked in June and October 2009 and July and October 2010. The highest species density was recorded in October. The maximum temperature was correlated positively with mean temperature and negatively with rainfall and relative humidity. Rainfall showed a positive correlation with mosquito abundance and species density. These data will be useful for understanding the transmission of arboviruses and human malaria in the region.

Blowflies (Diptera: Calliphoridae) are necrophagous flies with ecological, medical, veterinary, and forensic importance. These flies carry pathogens that they spread to animals and humans, and produce myiasis. Furthermore, they are useful tools in forensic science as indicators of postmortem intervals, and in biomedicine they are successfully used in larval therapy.This study aimed to assess the spatiotemporal variation of the blowflies community from the urban area of Sincelejo City, located in the Colombian Caribbean region. Samplings were conducted from May 2012 to April 2013, using Van Someren Rydon-traps baited with bovine meat, fish, and chicken that decomposed for 48 h. Six species were registered, Cochliomyia macellaria (Fabricius, 1775) was the most abundant, followed by Lucilia eximia (Wiedemann, 1819) and Chrysomya albiceps (Wiedemann, 1819), while Chloroprocta idioidea (Robineau-Desvoidy, 1830), Chrysomya megacephala (Fabricius, 1794), and Lucilia cuprina (Wiedemann, 1830) presented the least individuals.The blowflies community did not change significantly throughout the sampling months or between the periods of low or high rainfall, nor was it influenced by weather variables. Although the species composition was the same in all sampled localities, species abundances presented significant differences.The possible causes of this pattern are discussed herein. Sex ratios were female biased for all the recorded species, except C. idioidea. Our findings provide the first assessment of the blowflies community from the urban area of Sincelejo City.

The Asian tiger mosquito [Aedes albopictus (Skuse)] is an invasive species of public health importance that is currently expanding its range in the Northeast United States. Effective Ae. albopictus control depends on a thorough understanding of factors influencing their abundance, spatial distribution, and habitat preference. We conducted a series of container surveys for Ae. albopictus larvae/pupae over 2 yr across nine sites in neighborhoods along its invasive range in southern New York. Selected sites represented a gradient of percent impervious surface and median household income. Two hypotheses were tested: 1) Ae. albopictus larval/pupal abundance increases and spatial distribution becomes less clustered as site-level median income decreases and percent impervious surface increases because of increased larval habitat quality and availability; and 2) container-level characteristics are predictive of Ae. albopictus larval/pupal infestation across a range of sites. In 2016, neither median household income nor impervious surface predicted Ae. albopictus abundance. In 2017, sites with greater impervious surface were more heavily infested by some measures. In both years, Ae. albopictus larval/pupal spatial distribution as measured by K-function was more clustered in with greater median household income. Most container characteristics were either not predictive of Ae. albopictus or varied between years. Based on the variability of predictive container characteristics, we conclude that identification of key containers is not useful in this region. However, Ae. albopictus can be nonhomogenously distributed or abundant based on income level and impervious surface. Improved control of immatures should consider these regional predictors of Ae. albopictus populations.

Native and invasive container-inhabiting Aedes mosquitoes (Diptera: Culicidae) play important roles in the transmission of endemic and traveler-introduced arboviruses in the United States. In response to the emergence of Zika virus into the Americas, we surveyed the distribution of container Aedes spp. of public health importance within North Carolina during 2016 using ovitraps. A seasonal survey was conducted in 18 counties from the mountains to the coast to identify species incriminated in the transmission of chikungunya, dengue, La Crosse, yellow fever, and Zika viruses. Multiple local, state, and federal agencies participated in the study and submitted more than 3,600 ovistrips. Aedes albopictus (Skuse) (81.4%, n = 54,458) was the most common and widespread species found in this survey, followed by Aedes triseriatus (Say) (10.7%, n = 7,169) and Aedes japonicus (Theobald) (7.9%, n = 5,262). We did not find Aedes aegypti and rarely found Aedes hendersoni (Cockerell). We assessed broad-scale climatic and other factors and determined that longitude, elevation, rainfall, and temperature had significant effects on explaining the variation in presence, abundance, and phenology of container Aedes in North Carolina. However, much of the variation in these outcomes was not explained at this coarse scale and may benefit from finer-scale analyses. These efforts represent the largest ovitrap survey ever conducted in the state.

Knowledge about mosquito species diversity at tropical montane cloud forests (TMCFs) in Mesoamerica is scarce. Here, we present data on mosquito species richness from samples biweekly collected, from January to December 2017, in ovitraps installed in aTMCF patch at Vázquez de Coronado County, Costa Rica. Ovitraps were placed at 2.25, 1.50 and 0.75 m at 16 sampling points. During the study period we measured relative humidity and air temperature at each sampling point, and water temperature, volume and pH in each ovitrap. We collected a total of 431 mosquito larvae belonging to five taxonomic units, one identified to the genus level and four to the species level. The most common mosquito species was Culex bihaicolus Dyar & Nuñez Tovar (Diptera: Culicidae), which accounted for nearly 80% (n = 344) of the collected mosquitoes. Culex nigripalpus Theobald (Diptera: Culicidae) was the only medically important species we found and it was collected both in the dry (January to March) and rainy season (April to December). Over 95% (n = 411) of the mosquitoes were collected during the rainy season and 60% (n = 257) at 0.75 m. Among the environmental variables that we measured, only water volume and pH were significantly (P < 0.05) different between the dry and rainy season, the former increasing and the later decreasing during the rainy season. These results suggest that rainfall plays a major role regulating the phenology of the sampled mosquito species and highlight the need to screen for pathogens in Cx. nigripalpus at the study area.

Musca domestica L., the common housefly, is a very important mechanical vector of pathogens. Continuous exposure to pyrethroid insecticides has led to insecticide resistance in houseflies. Some mutations in the voltage-gated sodium channel gene (vgsc) reduce the binding affinity of pyrethroids target site insensitivity. We collected houseflies from the Urmia district of Northwestern Iran. Following DNA extraction, 580 bp regions of the vgsc known to contain knockdown resistance (kdr) mutations were amplified and sequenced using specific primers. The amplified region contained two exons (211-bp and 248-bp) and three introns. There were eight polymorphic sites between M. domestica insecticide-susceptible (MDU38813), super-kdr (NW_004774263) and aabys (KT897924) strains from GenBank in comparison with our sequences. Two amino acid substitutions were detected, N967Y (% polymorphism = 9.5%) and L1014H (% polymorphism = 4.7%) that can be associated with resistance. The common and previously reported mutations L1014F and M918T+L1014F were not detected. Diagnosis based on sequence analysis is useful for monitoring the frequency of pyrethroid resistance mutations, which will be helpful in avoiding overuse of this class of insecticides in house fly control.

Biorational mosquito larvicides based on microbial organisms and insect growth regulators (IGRs) have played a vital role in integrated mosquito control, particularly since the invasion of West Nile virus to the United States in 1999. Products that are formulated with technical powder of the bacterium, Bacillus sphaericus Neide (recently Lysinibacillus sphaericus Meyer and Neide), are among the ones that have been extensively applied to combat Culex and other mosquito species. Due to the simplicity of the binary toxins, resistance to this pesticide in laboratory and field populations of Culex pipiens L. complex has occurred globally since 1994. A Cx. pipiens population with a high level of resistance to B. sphaericus (VectoLex WDG) was identified in Salt Lake City, UT, in September 2016.The resistance ratios in this population were 20,780.0- and 23,926.9-fold at LC₅₀ and LC₉₀, respectively, when compared with a susceptible population of a laboratory reference colony of the same species. This B. sphaericus-resistant population remained mostly susceptible to other commonly used pesticides to control arthropods of public health and urban significance, including ones based on microbial organisms (Bacillus thuringiensis subsp. israelensis, spinosad, spinetoram, abamectin), IGRs (pyriproxyfen, methoprene, diflubenzuron, novaluron), organophosphate (temephos), neonicotinoid (imidacloprid), phenylpyrazole (fipronil), oxadiazine (indoxacarb), and pyrethroid (permethrin). Results are discussed according to the modes of action of the pesticides tested, and suggestions are made to manage B. sphaericus-resistant mosquito populations.

This study was conducted to monitor the susceptibility status of Aedes aegypti (Linnaeus) larvae in the Sunda Islands of Indonesia against various organophosphates and organochlorines. Larval bioassay was performed in accordance with the World Health Organization standard protocol. Field-collected and reference strains of Ae. aegypti larvae were tested against diagnostic doses of eight larvicides belonging to organophosphates and organochlorines, namely bromophos (0.050 mg/liter), chlopyrifos (0.002 mg/liter), fenitrothion (0.020 mg/liter), fenthion (0.025 mg/liter), malathion (0.125 mg/liter), temephos (0.012 mg/liter), DDT (0.012 mg/liter), and dieldrin (0.025 mg/liter). Mortality rates of larvae were recorded at 24-h posttreatment. This study showed that Ae. aegypti larvae from Padang, Samarinda, Manggarai Barat, and South Central Timor were susceptible to both fenitrothion and dieldrin (mortality rates ≥ 98%). About 6 out of 10 field strains of Ae. aegypti larvae were resistant (<80% mortality rates) against fenthion, whereas Ae. aegypti larvae from Kuningan, Samarinda, Sumba, and South Central Timor exhibited some degrees of resistance (mortality rates 80–98%). All field-collected Ae. aegypti larvae were resistant against diagnostic doses of chlorpyrifos, malathion, temephos, and DDT with mortality rates ranging from 0 to 74.67%. Continued insecticide susceptibility studies are essential to identify the efficacy of insecticides for an improved dengue vector control and to delay the development of insecticide resistance.

Phlebotomine sand flies are hematophagous insect vectors of the protozoan parasites of the genus Leishmania Ross, 1903 (Kinetoplastida: Trypanosomatida) that infect mammals, including humans, causing leishmaniasis. In Porto Alegre, Brazil, three autochthonous cases of human visceral leishmaniasis were reported in 2016 through 2017. We analyzed for the presence of Leishmania DNA in sand flies collected at the neighborhood of Agronomia, Porto Alegre, Brazil. Phlebotomine sand flies were collected at three sites from October 2014 to September 2015. Female sand flies were pooled in numbers from 1 to 20 depending upon species, locality, and date; all were screened for Leishmania infection by the amplification of the ITS1 region. In total 518 phlebotomine sand flies were collected: Psathyromyia lanei (Barretto and Coutinho, 1941) (Diptera: Psychodidae) (30.5%), Brumptomyia sp. (França and Parrrot, 1921) (Diptera: Psychodidae) (25.7%), Migonemyia migonei (Franca, 1920) (Diptera: Psychodidae) (21.4%), Pintomyia fischeri (Pinto, 1926) (Diptera: Psychodidae) (21.4%), and Nyssomyia neivai (Pinto, 1926) (Diptera: Psychodidae) (1.0%). Most sand flies were collected during the hot and rainy season from October 2014 to April 2015. Of the 113 pools analyzed, five pools of Pi. fischeri were PCR-positive with the amplicons possessing sequences similar (>95%) to that of Leishmania infantum Nicolle, 1908 (Kinetoplastida: Trypanosomatida). These results represent the first molecular detection of Le. infantum in Pi. fischeri. It is possible that Pi. fischeri is involved in the transmission cycle of Le. infantum in the studied area; however, further studies are needed to establish the true role of Pi. fischeri in the visceral leismaniasis cycle.

Rickettsia felis (Rickettsiales: Rickettsiaceae) is an emergent human pathogen that causes febrile illnesses in various parts of the world. This study describes the identification and growth characteristics of a R. felis-like organism (designated as Rickettsia sp. TH2014) cultured from Ctenocephalides orientis fleas in rural Malaysia. In this study, culturing of rickettsiae from filtered triturated flea lysates was performed in Aedes albopictus C6/36 cells. Cytopathic effects were observed from one of the samples 4 d post-inoculation. Electron microscopy revealed actively replicating intracytosolic coccobacillary organisms in the rickettsia-infected cells. Sequence analysis of amplified citrate synthase (gltA) gene fragment shows complete match of the rickettsia with Rickettsia sp. Rf31 in Southeast Asia, and ‘Candidatus Rickettsia senegalensis' strain PU01-02 in Africa. The whole-genome sequence of Rickettsia sp. TH2014 was determined and assembled. The estimated genome size and guanine + cytosine content of the rickettsia are 1.37 Mb and 32.9%, respectively. The high values of average nucleotide identity and tetra-nucleotide signature correlation index obtained from pairwise genome comparison study suggest the identification of the rickettsia as R. felis.The whole-genome single-nucleotide polymorphism analysis demonstrates close genetic relatedness of the rickettsia with R. felis and Rickettsia asemboensis. However, based on sequence analyses of rickettsial genes (16S rDNA, gltA, ompB, and sca4), Rickettsia sp. TH2014 is found to be distinct from R. felis and R. asemboensis.The sequence analyses reveal that Rickettsia sp.TH2014 is highly similar to ‘Ca. Rickettsia senegalensis' detected in fleas from Africa, Asia, and North America. Further investigation to provide insights on pathogenic potential and transmission dynamics of the rickettsia is warranted.

To test their virus susceptibility and capacity as transmission vectors, Japanese Culicoides species were artificially fed a mixture of Akabane virus (AKAV) and bovine blood, and virus recovery was attempted from infected midges 9–11 d post-exposure. Culicoides tainanus, C. punctatus, C. humeralis, C. jacobsoni, C. oxystoma, and C. asiana were found to be orally susceptible to AKAV. Virus titers in single infected midges of C. tainanus, C. oxystoma, C. punctatus, and C. jacobsoni ranged from 10^0.75 to 10^4.0 TCID₅₀ (tissue culture infectious dose). The titers in the infected C. oxystoma were significantly higher than those in the other infected species. Viral RNA was detected from both midges testing positive and those testing negative for infectious virus particles, but the viral RNA copies in the infectious virus-negative midges were significantly lower than those in the infectious virus-positive midges. Lower viral amplification, limited dissemination or both caused by tissue barriers might occur in infected midges from which infectious viruses were undetectable. A fully disseminated infection was developed in orally infected C. oxystoma and C. tainanus. This finding indicates their capacity to transmit AKAV, assuming that salivary gland barriers have limited effects on viral entry to and replication in salivary gland tissue. This result also suggests that the other orally susceptible species are potentially competent for AKAV transmission and would be considered active vectors of its spread.

Roads and highways can affect the spread of insect-borne diseases by limiting or amplifying the spatiotemporal distribution of vectors, pathogens, and hosts, which can, in turn, lead to the creation of a nidus of infection.The aim of this study was to compare the diversity (richness and abundance) of phlebotomine sand flies in household and forest edge environments found along two different segments of an Amazonian highway. Sampling was conducted along the northern and southern portions of highway BR-319, in Amazonas State, Brazil. At each sampling point, Hoover Pugedo traps were set in indoor and outdoor habitats, and at forests edges, and captures were made between 06:00 pm and 06:00 am. A total of 1,189 sand flies were captured and 48 species were identified. As expected, a greater number of species and individuals were captured in forest edge environments. Permutational Multivariate Analyses of Variance (PERMANOVA) and Permutational Analyses of Multivariate Dispersions (PERMDISP) analyses showed that sand fly fauna differed significantly among habitats, but no variance in species composition was observed between the two road segments. Some of the captured species were species that have been implicated as vectors of Leishmania spp. Ross, 1903 (Kinetoplastida: Trypanosomatidae).

High seropositivity to Rickettsia conorii and Rickettsia felis has been reported in Malaysian indigenous community living in settlements adjacent to forest areas. The current study was conducted to determine the type and distribution of rickettsiae in feeding and questing ticks that were collected from a forest reserve area at Kuala Lompat in Pahang, Malaysia. Using PCR assays targeting citrate synthase (gltA), outer membrane protein A (ompA) and B (ompB) genes, rickettsiae were detected from approximately one-third of 98 ticks (mainly Dermacentor and Haemaphysalis spp.) collected from the forest reserve. BLAST analysis reveals the predominance of Rickettsia sp. RF2125 in both feeding and questing ticks and Rickettsia sp. TCM1 in the questing ticks. Sequences exhibiting close genetic relationship with Rickettsia raoultii, Rickettsia tamurae, Rickettsia heilongjiangensis, and Rickettsia asiatica were also detected from the ticks. This study highlights the diversity of rickettsial species and potential tick vectors which may contribute to the high seropositivity observed among the local communities.

The mosquito Culex pipiens is the primary vector of Rift Valley fever, West Nile, encephalitis, and Zika viruses, and periodic lymphatic filariasis. Developing insecticide resistance in mosquitoes demands the development of new approaches to fight these diseases. Paratransgenesis and RNAi approaches by using engineered bacteria have been shown to reduce mosquito vector competence. Serratia-AS1 is a bacterium found in mosquitoes and was genetically modified for expression of antimalaria effector molecules that repress development of malaria parasites in mosquitoes. The aim of this study was to determine how a genetically marked Serratia strain expressing the mCherry fluorescent protein (mCherry-Serratia) affects the colonization potential, life span, blood feeding behavior, fecundity, and fertility of Cx. pipiens. mCherry-Serratia bacteria disseminated into larvae, pupae, and newly emerged adults and dramatically increased in numbers following a blood meal. The bacterium was transmitted to progeny, showing that it can extend horizontally, transstadially, and vertically through the mosquito population.The presence of mCherry-Serratia did not affect blood feeding behavior, survival rate, fecundity, and fertility of Culex mosquitoes. This is the first study to evaluate the effects of an engineered bacteria on the fitness of Cx. pipiens. Although challenges remain, such as producing engineered bacteria to secrete anti-pathogens associated with Cx. pipiens, introducing such bacteria into mosquito populations, our findings of minimal fitness cost caused by Serratia-AS1 bode well for the development of paratransgenesis and RNAi approaches.

In Tunisia, 17 phlebotomine sand fly species are reported, belonging to genera Phlebotomus and Sergentomyia. However, subsequent to faunal studies and outbreaks of leishmaniasis in different regions of the country, unrecognized sand fly species could exist. Indeed, we report in this study the presence of Sergentomyia (Parrotomyia) lewisi Parrot 1948. A brief collection of phlebotomine sand flies was undertaken in August 2016 in Khbina locality, in Sidi Bouzid governorate situated in CentralTunisia, which constitutes an old focus of Zoonotic Cutaneous Leishmaniasis. Sand flies were collected, using CDC light traps that were placed overnight, in different biotopes (inside habitations, outdoors, and within animal shelters). Specimens were collected and morphologically identified. Measurements were taken with an ocular micrometer.Two female specimens of the species Se. (Parrotomyia) lewisi were collected. One specimen is here described and measured. A comparison of its taxonomic characters to the holotype from Sudan and a specimen from Algeria is also presented. This species was until now only reported from Sudan, Ethiopia, Algeria, and Morocco. It is here described for the first time inTunisia, which raises theTunisian sand fly fauna to seven subgenera and 18 species.

The Transphlebotomus subgenus has been drawing attention in last decade due to the uncertain vector capacity and cryptic life history of the group. During a 2015 entomological survey, Phlebotomine flies were collected in western locations of Slovenia, with a total of 22 specimens of the Transphlebotomus subgenus collected. In addition to morphological identifications, we confirmed the presence of Phlebotomus mascittii Grassi, 1908 (Diptera: Psychodidae) by nucleotide sequences of the mitochondrial cytochrome b gene. During the study, we found that P. mascittii is most abundant in the area of coastal–karst region of Slovenia, while in rare occasions can also be collected in Northeastern part of the country, near the Austrian border. Vector competence of P. mascitii, like all other member species of the Transphlebotomus subgenus, is unclear, but potential to transmit Leishmania (Kinetoplastida: Trypanosomatidae) pathogens or phleboviruses is strongly suspected.

Haemaphysalis flava (Acari: Ixodidae) harbors pathogenic microorganisms and transfers these to hosts during blood feeding. Proteomic analysis in the midgut contents of H. flava detected glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and contig 1683 was retrieved as a GAPDH gene fragment by searching our previous transcriptomic library. In the study, the 5′ and 3′ ends of contig 1683 were cloned by rapid amplification of cDNA ends (RACE) and a full length, 1340 bp cDNA of Hf-GAPDH was obtained. The open-reading frame had 999 bp and coded for 333 amino acids. Hf-GAPDH was predicted to have an N-terminal NAD binding domain and a C-terminal glyceraldehyde dehydrogenase catalytic domain. The molecular structure of Hf-GAPDH was analyzed and the evolutionary relationship also established. The GAPDH protein sequence was conserved among ticks. The expression pattern of Hf-GAPDH, analyzed by real-time PCR, significantly differed among life phases, feeding stages, and tissues. As the ticks grew, the expression level of Hf-GAPDH was up-regulated. The expression levels of Hf-GAPDH in salivary glands and midguts from half-engorged ticks were lower than the same tissues from engorged ticks. This study will provide reference data for the follow-up verification of the GAPDH-related function and the feasibility as a potential anti-tick vaccine.

Aedes aegypti Linnaeus (Diptera: Culicidae) is a primary vector of multiple arboviruses including Zika, dengue, chikungunya, and yellow fever. After incorporating BG lure or BG lure + octenol, the CDC's Autocidal Gravid Ovitrap (AGO) was more effective at collecting and controlling host-seeking female Ae. aegypti than gravid female mosquitoes. The addition of octenol to the AGO did not increase the number of female mosquitoes captured by the AGO, compared with the AGO alone. The AGO baited with the BG lure captured a significant number of host-seeking female Ae. aegypti.This finding indicates that the combination of AGO with BG lure could enable the trap to control female Ae. aegypti mosquitoes representing either host-seeking or gravid physiological states.

Tick-borne relapsing fever (TBRF) is a potentially serious vector-borne disease endemic to the western United States. Vector surveillance is compromised by the nidicolous life history of the three Ornithodoros species that transmit TBRF to people in this region. Large-scale stationary trapping methods were developed to survey a wide geographical range of Ornithodoros spp. which are known to vector relapsing fever Borrelia spp. in California. Ninety-six Ornithodoros parkeri were collected from four locations in the foothills of Fresno and Madera Counties. Two of these O. parkeri nymphs were PCR positive for Borrelia parkeri, and their collection at a popular recreation site increases the public health concern.

Western equine encephalitis (WEE) was once prevalent and routinely isolated from mosquitoes in Colorado; however, isolations of Western equine encephalitis virus (WEEV) have not been reported from mosquito pools since the early 1990s.The objective of the present study was to test pools of Culex tarsalis (Coquillett) mosquitoes sampled from Weld County, CO, in 2016 for evidence of WEEV infection. Over 7,000 mosquitoes were tested, but none were positive for WEEV RNA.These data indicate that WEEV either was not circulating enzootically in Northern Colorado, was very rare, and would require much more extensive mosquito sampling to detect, or was heterogeneously distributed spatially and temporally and happened to not be present in the area sampled during 2016. Even though the reported incidence of WEE remains null, screening for WEEV viral RNA in mosquito vectors offers forewarning toward the detection and prevention of future outbreaks.